Mortality and decreased weight gain resulting from infection and disease in dairy calves is a problem within the dairy industry. Colostrum is the sole source of maternal immunity for the calf, having a substantial impact on health and survival. To date, colostrum quality is determined by concentration of antibodies. Colostrum also contains proteins and cells, which may enhance immune development in the neonate. Our goals were to determine the impact of colostral immune cells on (1) immune status during the first month of life and (2) immune development over time. To determine the impact of adoptively transferred colostral immune cells, calves were fed either whole colostrum (WC) or cell-free colostrum (CFC) at birth. During the first month of life, calves fed CFC had decreased numbers of CD4+ T cells when compared to WC-fed calves. However, CFC-fed calves had a greater percentage of monocytes during the first month of life. To determine the influence of colostral immune cells on immune development, cellular blood parameters were measured in response to two series of vaccinations (A and B). After vaccination series A, CFC-fed calves had decreased numbers of B cells when compared to WC-fed calves. After vaccination series B, CFC-fed calves had decreased levels of interleukin-2 gene expression and numbers of CD4+ and gamma delta T cells when compared to WC-fed calves. This study demonstrates that colostral immune cells impact immune status and development in dairy calves. / Master of Science

O objetivo geral deste trabalho foi avaliar a influência dos leucócitos do colostro bovino na imunidade inata, desenvolvimento da microbiota intestinal e ocorrência de diarreias em bezerras Holandesas recém-nascidas. Para isso, 20 bezerras Holandesas foram acompanhadas nos seguintes momentos: antes da mamada do colostro (D0); 1-2 (D2); 7 (D7); 14 (D14); 21 (D21) e 28 (D28) dias após nascimento e foram distribuídas em dois grupos experimentais: grupo COL+ recebeu colostro fresco (4L) proveniente de suas respectivas mães; e grupo COL- recebeu colostro congelado e acelular (4L) de vacas doadoras. Capítulo 1 O objetivo deste capítulo foi avaliar a presença e influência dos leucócitos do colostro na colonização do trato gastrintestinal. Para isso, amostras de colostro foram semeadas em TSA e MacConkey para CBT e CCT; e amostras de fezes em agar sangue, MacConkey e Salmonella-Shiguella. Bezerras apresentaram maior frequência de diarreia no D14 (COL+=78%; COL-=60%). O grupo COL- apresentou maior CBT e CCT/mL (3,93x106 ufc/mL e 3,01x105 ufc/mL) em relação ao COL+ (0,94x106 ufc/mL e 0,78x105ufc/mL). Sobre as espécies bacterianas isoladas, as que foram mais frequentes no COL+ foram Proteus mirabilis (29,91%), Escherichia coli (28,04%), Citrobacter freundii (5,61%) e Staphylococcus spp (5,61%), no COL- foram Escherichia coli (28,70%), Proteus mirabilis (27,78%), Klebsiella pneumoniae (11,32%) e Morganella morganii (5,66%). O momento com maior isolamento em ambos os grupos foi o D2 (COL+ = 26 cepas; COL- = 27 cepas), período em que iniciou a diarreia nas bezerras. Não foi possível detectar diferenças entre as frequências de microrganismos entre os grupos COL+ e COL-. A administração de colostro COL+ e COL- não influenciou na proporção de bactérias aeróbias presentes nas fezes e na ocorrência de diarreia das bezerras durante o período neonatal. Capítulo 2 O objetivo deste capítulo foi avaliar a influência dos leucócitos do colostro na resposta imune inata em bezerras Holandesas. Os animais foram submetidos ao exame clínico, seguido da colheita das amostras sanguíneas para realização das provas laboratoriais. As concentrações de cortisol sérico e haptoglobina não apresentaram diferenças estatísticas, enquanto a concentração de ferro foi diferente entre os grupos no D7. No leucograma não foi detectada diferença para os leucócitos totais e neutrófilos absolutos, mas houve tendência entre os grupos no D7 para os valores de neutrófilos relativos. O marcador CH138+CD62L+ não apresentou diferença entre os grupos, apesar da maior expressão ter sido detectada no COL-. A expressão de CH138+CD62L- foi maior no COL+ no D14. A fagocitose e produção de H2O2 pelos neutrófilos sanguíneos apresentou perfil semelhante em relação ao S. aureus e E. coli entre os grupos, apesar do COL- apresentar maior intensidade de fagocitose do D7 ao D14. Houve tendência para o aumento da proporção de granulócitos liberando H2O2 basal no D14 para o COL-, entretanto o COL + apresentou tendência para maior intensidade da fagocitose no D21. A proporção de granulócitos (%), estimulados com S. aureus, liberando H2O2 foi maior no COL+ em D7, enquanto o estímulo com a E. coli não resultou em diferenças entre os grupos. A intensidade de fluorescência foi maior e gradual no COL+ quando as células foram estimuladas, porém não foram encontradas diferenças estatísticas. As células do colostro influenciaram no perfil sanitário apresentado pelas bezerras, observando-se maior intensidade de diarreias, menores teores de ferro sérico e anemias no grupo que recebeu colostro congelado. A migração dos granulócitos sanguíneos foi mais rápida e intensa no COL+ em relação ao COL-, após exposição natural aos patógenos causadores de diarreia. O índice de fagocitose (%) dos granulócitos apresentou semelhança entre os grupos, entretanto a intensidade da fluorescência foi mais intensa no COL-. Em contrapartida, o COL+ demonstrou maior habilidade em produzir H2O2 / The aim of this study was to evaluate the influence of bovine colostrum leukocytes in innate immunity, in the development of the intestinal microbiota and the occurrence of diarrhea in newborn Holstein calves. For this, 20 Holstein calves were followed in the following moments: before colostrum intake (D0); 1-2 (D2); 7 (D7); 14 (D14); 21 (D21) and 28 (D28) days after birth. They were divided into two experimental groups: COL+ group, which received fresh colostrum (4L) from their mothers; and COL-group which received frozen and acellular colostrum (4L) of donor cows. Chapter 1 -The aim of this chapter was to evaluate the presence and influence of colostrum leukocytes in the colonization of the gastrointestinal tract. For this, colostrum samples were seeded in TSA and MacConkey for TPC and TCC; and stool samples on blood agar, MacConkey and Salmonella-Shiguella. Calves presented higher frequency of diarrhea in D14 (COL+ = 78%; COL- = 60%). The COL- group showed higher TPC and TCC/mL (3,93x106 cfu/mL and 3,01x105 cfu/mL) compared to COL+ (0,94x106 cfu/mL and 0,78x105 cfu/mL). Regarding the isolated bacterial species, those that were more frequent in the COL+ were Proteus mirabilis (29.91%), Escherichia coli (28.04%), Citrobacter freundii (5.61%) and Staphylococcus spp (5.61%); in the COL-were Escherichia coli (28.70%), Proteus mirabilis (27.78%), Klebsiella pneumoniae (11.32%) and Morganella morganii (5.66%). The moment with more insolation in both groups was the D2 (COL+ = 26 strains; COL- = 27 strains), period in which started the diarrhea in calves. It was not possible to detect differences between the frequencies of microorganisms between the COL+ and COL- groups. The COL+ and COL- colostrum management did not influence the proportion of aerobic bacteria present in feces and the occurrence of diarrhea in calves during the neonatal period. Chapter 2 The aim of this chapter was to evaluate the influence of colostrum leukocytes in the innate immune response in Holstein calves. The animals were submitted to clinical examination, followed by blood samples collection to perform laboratory tests. The serum cortisol and haptoglobin concentrations did not show statistical differences while the iron concentration was different between groups at D7. The leukogram did not show difference to total leukocytes and absolute neutrophils, but there was a trend between groups at D7 for relative values of neutrophils. The CH138+CD62L+ marker showed no difference between groups, despite an increased expression was detected in COL-. The expression of CH138+CD62L- was greater in COL+ at D14. The phagocytosis and production of H2O2 by blood neutrophils showed similar profile in relation to S. aureus and E. coli between groups, although COL- presented greater phagocytosis intensity from D7 to D14. There was a trend for the increase of granulocyte proportion, releasing basal H2O2 at D14 for COL-, however COL+ presented trend to a higher intensity of phagocytosis at D21. The proportion of granulocytes (%) stimulated with S. aureus releasing H2O2 was greater in COL+ at D7, while stimulation with E. coli resulted in no differences between the groups. The fluorescence intensity was higher and gradual in COL+ when cells were stimulated; however, it was not detected statistical differences. The colostrum cells influenced in the health profile presented by the calves, with a higher intensity of diarrhea, lower levels of serum iron and anemia in the group that received frozen colostrum. The migration of blood granulocytes was faster and more intense in COL+ in relation to COL- after natural exposure to pathogens that cause diarrhea. The phagocytosis rate (%) of granulocytes showed similarity between groups, although the fluorescence intensity of COL- was more intense. In contrast, the COL+ demonstrated greater ability in producing H2O2

O objetivo geral deste trabalho foi avaliar a influência dos leucócitos do colostro bovino na imunidade inata, desenvolvimento da microbiota intestinal e ocorrência de diarreias em bezerras Holandesas recém-nascidas. Para isso, 20 bezerras Holandesas foram acompanhadas nos seguintes momentos: antes da mamada do colostro (D0); 1-2 (D2); 7 (D7); 14 (D14); 21 (D21) e 28 (D28) dias após nascimento e foram distribuídas em dois grupos experimentais: grupo COL+ recebeu colostro fresco (4L) proveniente de suas respectivas mães; e grupo COL- recebeu colostro congelado e acelular (4L) de vacas doadoras. Capítulo 1 O objetivo deste capítulo foi avaliar a presença e influência dos leucócitos do colostro na colonização do trato gastrintestinal. Para isso, amostras de colostro foram semeadas em TSA e MacConkey para CBT e CCT; e amostras de fezes em agar sangue, MacConkey e Salmonella-Shiguella. Bezerras apresentaram maior frequência de diarreia no D14 (COL+=78%; COL-=60%). O grupo COL- apresentou maior CBT e CCT/mL (3,93x106 ufc/mL e 3,01x105 ufc/mL) em relação ao COL+ (0,94x106 ufc/mL e 0,78x105ufc/mL). Sobre as espécies bacterianas isoladas, as que foram mais frequentes no COL+ foram Proteus mirabilis (29,91%), Escherichia coli (28,04%), Citrobacter freundii (5,61%) e Staphylococcus spp (5,61%), no COL- foram Escherichia coli (28,70%), Proteus mirabilis (27,78%), Klebsiella pneumoniae (11,32%) e Morganella morganii (5,66%). O momento com maior isolamento em ambos os grupos foi o D2 (COL+ = 26 cepas; COL- = 27 cepas), período em que iniciou a diarreia nas bezerras. Não foi possível detectar diferenças entre as frequências de microrganismos entre os grupos COL+ e COL-. A administração de colostro COL+ e COL- não influenciou na proporção de bactérias aeróbias presentes nas fezes e na ocorrência de diarreia das bezerras durante o período neonatal. Capítulo 2 O objetivo deste capítulo foi avaliar a influência dos leucócitos do colostro na resposta imune inata em bezerras Holandesas. Os animais foram submetidos ao exame clínico, seguido da colheita das amostras sanguíneas para realização das provas laboratoriais. As concentrações de cortisol sérico e haptoglobina não apresentaram diferenças estatísticas, enquanto a concentração de ferro foi diferente entre os grupos no D7. No leucograma não foi detectada diferença para os leucócitos totais e neutrófilos absolutos, mas houve tendência entre os grupos no D7 para os valores de neutrófilos relativos. O marcador CH138+CD62L+ não apresentou diferença entre os grupos, apesar da maior expressão ter sido detectada no COL-. A expressão de CH138+CD62L- foi maior no COL+ no D14. A fagocitose e produção de H2O2 pelos neutrófilos sanguíneos apresentou perfil semelhante em relação ao S. aureus e E. coli entre os grupos, apesar do COL- apresentar maior intensidade de fagocitose do D7 ao D14. Houve tendência para o aumento da proporção de granulócitos liberando H2O2 basal no D14 para o COL-, entretanto o COL + apresentou tendência para maior intensidade da fagocitose no D21. A proporção de granulócitos (%), estimulados com S. aureus, liberando H2O2 foi maior no COL+ em D7, enquanto o estímulo com a E. coli não resultou em diferenças entre os grupos. A intensidade de fluorescência foi maior e gradual no COL+ quando as células foram estimuladas, porém não foram encontradas diferenças estatísticas. As células do colostro influenciaram no perfil sanitário apresentado pelas bezerras, observando-se maior intensidade de diarreias, menores teores de ferro sérico e anemias no grupo que recebeu colostro congelado. A migração dos granulócitos sanguíneos foi mais rápida e intensa no COL+ em relação ao COL-, após exposição natural aos patógenos causadores de diarreia. O índice de fagocitose (%) dos granulócitos apresentou semelhança entre os grupos, entretanto a intensidade da fluorescência foi mais intensa no COL-. Em contrapartida, o COL+ demonstrou maior habilidade em produzir H2O2 / The aim of this study was to evaluate the influence of bovine colostrum leukocytes in innate immunity, in the development of the intestinal microbiota and the occurrence of diarrhea in newborn Holstein calves. For this, 20 Holstein calves were followed in the following moments: before colostrum intake (D0); 1-2 (D2); 7 (D7); 14 (D14); 21 (D21) and 28 (D28) days after birth. They were divided into two experimental groups: COL+ group, which received fresh colostrum (4L) from their mothers; and COL-group which received frozen and acellular colostrum (4L) of donor cows. Chapter 1 -The aim of this chapter was to evaluate the presence and influence of colostrum leukocytes in the colonization of the gastrointestinal tract. For this, colostrum samples were seeded in TSA and MacConkey for TPC and TCC; and stool samples on blood agar, MacConkey and Salmonella-Shiguella. Calves presented higher frequency of diarrhea in D14 (COL+ = 78%; COL- = 60%). The COL- group showed higher TPC and TCC/mL (3,93x106 cfu/mL and 3,01x105 cfu/mL) compared to COL+ (0,94x106 cfu/mL and 0,78x105 cfu/mL). Regarding the isolated bacterial species, those that were more frequent in the COL+ were Proteus mirabilis (29.91%), Escherichia coli (28.04%), Citrobacter freundii (5.61%) and Staphylococcus spp (5.61%); in the COL-were Escherichia coli (28.70%), Proteus mirabilis (27.78%), Klebsiella pneumoniae (11.32%) and Morganella morganii (5.66%). The moment with more insolation in both groups was the D2 (COL+ = 26 strains; COL- = 27 strains), period in which started the diarrhea in calves. It was not possible to detect differences between the frequencies of microorganisms between the COL+ and COL- groups. The COL+ and COL- colostrum management did not influence the proportion of aerobic bacteria present in feces and the occurrence of diarrhea in calves during the neonatal period. Chapter 2 The aim of this chapter was to evaluate the influence of colostrum leukocytes in the innate immune response in Holstein calves. The animals were submitted to clinical examination, followed by blood samples collection to perform laboratory tests. The serum cortisol and haptoglobin concentrations did not show statistical differences while the iron concentration was different between groups at D7. The leukogram did not show difference to total leukocytes and absolute neutrophils, but there was a trend between groups at D7 for relative values of neutrophils. The CH138+CD62L+ marker showed no difference between groups, despite an increased expression was detected in COL-. The expression of CH138+CD62L- was greater in COL+ at D14. The phagocytosis and production of H2O2 by blood neutrophils showed similar profile in relation to S. aureus and E. coli between groups, although COL- presented greater phagocytosis intensity from D7 to D14. There was a trend for the increase of granulocyte proportion, releasing basal H2O2 at D14 for COL-, however COL+ presented trend to a higher intensity of phagocytosis at D21. The proportion of granulocytes (%) stimulated with S. aureus releasing H2O2 was greater in COL+ at D7, while stimulation with E. coli resulted in no differences between the groups. The fluorescence intensity was higher and gradual in COL+ when cells were stimulated; however, it was not detected statistical differences. The colostrum cells influenced in the health profile presented by the calves, with a higher intensity of diarrhea, lower levels of serum iron and anemia in the group that received frozen colostrum. The migration of blood granulocytes was faster and more intense in COL+ in relation to COL- after natural exposure to pathogens that cause diarrhea. The phagocytosis rate (%) of granulocytes showed similarity between groups, although the fluorescence intensity of COL- was more intense. In contrast, the COL+ demonstrated greater ability in producing H2O2

Environ 20% des chiots nés vivants meurent au cours des trois premières semaines de vie. Cette mortalité néonatale est ainsi responsable de pertes économiques importantes pour les éleveurs. Néanmoins, les déterminants immunitaires et nutritionnels de ces cas de mortalité sont très mal connus dans l'espèce canine. L'objectif de ce travail était donc d'identifier les facteurs de risque de mortalité néonatale chez le chiot, dont en particulier l'importance de la prise colostrale pour la survie. La première partie des résultats a mis en évidence une association forte entre la croissance précoce (sur les deux premiers jours de vie) et les pertes néonatales. Cette croissance précoce reflétant directement la prise colostrale, la suite des travaux s'est ensuite intéressée aux différents apports colostraux. Le transfert d'immunité passive affecte le taux de mortalité, une concentration sérique en immunoglobulines G à l'âge de 2 jours inférieure à 2,3 g/l étant associée à un risque de mortalité néonatale plus élevé et caractérisant un déficit de transfert. Un résultat similaire a été obtenu avec l'étude de l'immunité spécifique dirigée contre le parvovirus canin de type 2 : les chiots ayant acquis les titres en anticorps les plus faibles à deux jours d'âge séroconvertissent et excrètent du virus significativement plus tôt que les chiots avec de plus forts titres d'anticorps maternels. L'apport énergétique colostral était également associé aux chances de survie : les chiots présentant une glycémie à 24 heures de vie inférieure à 0,92 g/l ayant un plus fort risque de mortalité. Outre la forte relation entre survie et prise colostrale (apportant immunoglobulines et énergie), notre travail montre également l'impact du poids et de la vitalité à la naissance (évalué par le score Apgar). Par ailleurs, la qualité immunologique du colostrum (évaluée par sa concentration en IgG), bien que non directement liée aux chances de survie, s'est montrée très variable entre les chiennes, aggravant probablement le risque de déficit de transfert de l'immunité passive chez certains chiots. Tous ces travaux contribuent à une meilleure connaissance des facteurs de risque de mortalité qu'il est important de contrôler en élevage. Ils révèlent le rôle crucial de la croissance foetale, du déroulement de la mise-bas et de l'ingestion colostrale. Une pesée des chiots au cours des premiers jours de vie peut être a minima conseillée aux éleveurs pour identifier les chiots à risque et auxquels des soins plus attentifs devront être apportés. / Neonatal mortality in dogs (within the first three weeks after birth) accounts on average for 20% of puppies born alive, being thus responsible for a great economic loss to dog breeders. However, immunological and nutritional determinants of puppies survival are poorly described. This dissertation thus investigated the risk factors of neonatal mortality in dogs, and in particular the importance of colostrum intake for survival. The first part of results revealed a strong association between early growth rate (during the first two days of life) and neonatal losses. As early growth rate reflects the colostrum intake, the role of colostrum was then addressed. Passive immune transfer was shown to affect mortality rate, with serum IgG concentration at two days of age lower than 2.3 g/L being characterized as in deficit of maternal immunoglobulins. Similar lack of immunoprotection was observed for a specific canine pathogen (canine parvovirus type 2), as puppies with low antibody titers at two days of age seroconverted or underwent parvovirus infection significantly earlier than puppies with higher titers. Energy intake, evaluated via blood glucose concentration at 24h of life, was also found associated with survival: puppies with low glucose concentration (≤ 92 mg/dl) were found at higher risk of death. Besides the strong relationship between colostrum ingestion, providing passive immunity and energy, the impact of birth weight and vitality at birth (evaluated via Apgar score) on puppies’ survival was also evidenced. Colostrum immune quality (evaluated via IgG concentration), although not directly linked with the risk of neonatal death, was found of great variability, most probably putting some puppies at a risk of passive immune deficit. The present study contributed to the knowledge about the risk factors of mortality to be controlled in breeding kennels. Results provided in this work revealed the crucial role of the fetal growth, course of parturition and intake of the colostrum for the newborn dog. Regular weighing of newborns can be advised as a practical application for dog breeders, as it allows to identify puppies at a higher risk of death and to provide puppies with additional nursing and veterinary care.

No description available.

To determine the antigenic relatedness of the different developmental stages of Cryptosporidium parvum, monoclonal IgG3 antibody (mAb), Cmg-3, was produced by immunizing mice with partially purified merozoites. The monoclonal Cmg-3 reacted with a 3.5 kDa antigen of sporozoites in western blots and appeared to react with cell surface antigens of air-dried merozoites and sporozoites using immunofluorescence (IF). Additional mAbs, C6B6 (IgG1) and C4A1 (IgM), which react with a 20 kDa and multiple sporozoite antigens, respectively, were employed for immunoelectron microscopic studies with Cmg-3. These mAbs showed similar (surface/cytoplasmic) immunoelectron microscopic colloidal gold labeling patterns with all C. parvum life cycle stages. The three mAbs were also examined for potential modulation of cryptosporidial infections in vivo by daily oral mAb administration to oocyst-inoculated neonatal mice. Monoclonal-treated neonatal mice were sacrificed four and eight days post infection (pi). Differences in infection rates were observed among the treatment groups (p < .05). Suckling mice treated daily with orally administered mixtures of mAbs (ascitic fluids) showed significantly reduced parasite loads compared to control mice at four and eight days pi, while suckling mice receiving mAb Cmg-3 alone showed significant differences only at four days pi. Passive transfer of immunity using hyperimmune bovine colostrum was performed to determine the therapeutic and prophylactic efficacy of daily oral administration of anti-C. parvum antibody on the manifestation of cryptosporidial disease in neonatal mice as a model for treating cryptosporidiosis in immunocompromised patients. Hyperimmune colostrum was found to provide therapeutic and prophylactic efficacy against cryptosporidiosis in neonatal mice. Significantly fewer (p < 0.05) stages of C. parvum were found in mice that received hyperimmune skim colostrum (HSC) or hyperimmune original colostrum (HC) than in those treated with control colostrum (CC) or saline. Using IF, antigen-specific IgG in HSC and HC to C. parvum was 35 times greater than that of CC. There was no significant difference between groups treated with HSC or HC (p < .05), which suggests that the immunoglobulins, other biologically active factors such as cytokines, or both, might be active factors of immunity against cryptosporidiosis.

No description available.

Existem fortes evidências de que a supressão da hipersensibilidade nos recém nascidos pode ser mediada pela transferência de anticorpos maternos, dependendo de sua concentração e especificidade, no entanto carece estudos sobre a eficácia em humanos. Realizamos este estudo a fim de caracterizar qualitativa e quantitativamente a transmissão de anticorpos direcionados ao principal alérgeno da poeira domiciliar (Der p) via placenta e colostro materno, assim como investigar o efeito da sensibilização materna ao Der p na transferência passiva destes anticorpos. Para tais objetivos, analisamos amostras de sangue materno, cordão umbilical e colostro de puérperas sensibilizadas. Demonstramos pela primeira vez que S-IgA anti-Der p pode ser transferida ao lactente em concentrações bastante variáveis e com alto índice de avidez, independente da sensibilização materna ao mesmo ácaro. Demonstramos também que o nível de IgG específica ao Der p é mais elevado em recém nascidos de mães sensibilizadas quando comparado aos de mães controle não sensibilizadas. Já a avidez específica da IgG anti-Der p foi muito semelhante entre as amostras pareadas de cordão umbilical e soro materno, assim como em amostras do grupo estudo e grupo controle. / It is known that the incidence of allergic disease has been rising very fast in the last decade and nowadays affects thousands of children worldwide. For this reason, it is of great interest that efficient strategies of prevention of atopy should be applied in the first years of life or even before birth. The are strong evidences that the suppression of hypersensitivity in newborn can be mediated by the transference of maternal antibodies, depending on their concentration and specificity, however still little is known about the mechanisms involving, in special in humans. We made this study aiming to characterize qualitatively and quantitatively the antibodies transmission directed to the main home dust allergen (Dermatophagoides pteronyssinus; Der p) through placental transference and maternal breastfeeding as well as to investigate the maternal sensitizing effect against to Der p in passive transference of these antibodies. For those objectives, we quantified by ELISA, IgG anti-Der p in paired samples of maternal blood and umbilical cord and anti-Der p S-IgA in colostrums of sensitized mother (n=13) and not sensitized (n=26); and we analyzed the functional activity of the same antibodies by avidity assays. The sensibility was determined in maternal sera by specific RAST (Cap System® Pharmacia). We show by the first time that anti-Der p S-IgA is transferred to the infant in very variable concentrations and with high levels of avidity, but is not dependent of maternal sensitization. We believe that breastfeeding is important, because it supplies S-IgA with the capacity to neutralize in a specific manner and block the entrance of Der p through mucosa, in infant of RAST+ mothers as well as of RAST-. We also demonstrate that total and specific to Der p IgG levels are more elevated in newborns of sensitized mothers when compared to of those of control mothers and non-sensitized indicating that the maternal sensitizing can influence the fetal immune response. In the other hand, the specific avidity of anti-Der p IgG was very similar between paired samples of umbilical cord and maternal sera, as well as in samples of study group and control group, suggesting thar the avidity index of IgG does not influence on placental transfer of specific antibodies. Once that the maternal antibody transference represent a important mechanism for immunomodulation of allergic response, we expect that a better understanding of the influence of maternal sensitivity on passive transfer of specific antibodies to babies will contribute with advances in the elaboration of adequate strategies of prevention of allergic sensitivity with more efficient therapeutic results.

O objetivo deste trabalho foi identificar o padrão de vigor e status metabólico através da determinação da frequência dos sinais objetivos e metabólitos sanguíneos após o nascimento, possivelmente associados à transferência de imunidade passiva, além de determinar o melhor horário para o uso do refratômetro de Brix na avaliação da transferência de imunidade passiva em bezerros neonatos após a ingestão de colostro. No primeiro experimento foram utilizados 46 bezerros neonatos de ambos os sexos, nascidos entre setembro de 2013 e julho de 2014. A avaliação do vigor dos animais foi realizada entre 15 e 30 minutos após o nascimento com o auxílio da escala APGAR modificada pela Universidade de Guelph. A colheita de sangue para análise de metabólitos seguiu os horários de 0h (antes da ingestão), 1h, 2h, 4h, 6h, 12h, 24h, 48h contadas a partir da ingestão do colostro. Os parâmetros sanguíneos foram determinados por kit enzimático específico para cada parâmetro em Sistema Automático para Bioquímica. Todos os animais utilizados neste estudo foram oriundos de partos eutócicos e apresentaram, de acordo com a escala APGAR, vigor satisfatório. As concentrações de metabólicos sanguíneos dos neonatos mostraram que o consumo de colostro aumentou a disponibilidade de energia e a concentração de frações proteicas, comumente utilizadas como indicativos da transferência de imunidade passiva. No segundo experimento foram utilizados 47 bezerros neonatos de ambos os sexos, nascidos entre março e julho de 2014. O colostro foi ordenhado após o parto e a qualidade determinada com auxílio do colostrômetro e do refratômetro digital. A colheita do sangue para a análise dos parâmetros sanguíneos seguiu os horários de 0h (antes da ingestão), 1h, 2h, 4h, 6h, 12h, 24h, 48h, 72h, 96 e 120h, contadas a partir da ingestão do colostro. A concentração de proteína total foi determinada por refratômetro de Brix e por kit enzimático em Sistema Automático para Bioquímica. Os demais parâmetros sanguíneos foram determinados por kit enzimático específico para cada parâmetro em Sistema Automático para Bioquímica. A contagem global e diferenciada das células sanguíneas foi realizada nos tempos 0, 12 e 24h após o fornecimento do colostro. O colostro se manteve na faixa de boa qualidade, segundo os critérios de classificação de medidas pelo colostrômetro e refratômetro digital de Brix. O consumo de colostro foi o principal fator de variação dos parâmetros estudados neste trabalho. A avaliação da transferência de imunidade passiva em bezerros recém-nascidos deve ser realizada por volta das 24 horas após a ingestão do colostro, pois neste momento a absorção de macromoléculas colostrais já está encerrada e a PT alcança estabilidade. A partir deste período, não é possível determinar se as frações proteicas avaliadas são de origem colostral ou endógena, o que não garante avaliação segura sobre a transferência de imunidade passiva. / The objective of this study was to identify the vigor and metabolic status through the determination of objective signs and blood metabolites just after birth of dairy calves, possibly associated with the passive immunity transfer. Also, determine the best time after colostrum intake to estimate the passive immunity transfer using a Brix refractometer. The first trial evaluated 46 newborn calves, males and females, born from September 2014 to July 2014. Animals\' vigor evaluations were carried out between 15 and 30 minutes after birth using the APGAR scale modified by the University of Guelph. The blood sample collection for metabolites analysis were done at 0h (before intake), 1h, 2h, 4h, 6h, 12h, 24h, and 48h after colostrum intake. Blood parameters were analyzed by enzymatic kits using an automatic biochemical system. All animals were born from eutocic births and presented, according to the APGAR scale, satisfactory vigor. The newborns metabolites blood concentrations revealed that colostrum intake increased the energy availability and the concentration of protein fractions, commonly used as indicative of passive immunity transfer. For the second trial 47 newborn calves of both sexes, born from March and July 2014, were used. The colostrum was milked after parturition and quality was determined using a colostrometer and digital Brix refractometer. Blood samples were taken from calves at 0h (before intake), 1h, 2h, 4h, 6h, 12h, 24h, 48h, 72h, 120h and 96, after colostrum intake. The total protein (TP) concentration was measured by a Brix refractometer and an enzymatic kit using an automatic biochemical system. Other blood parameters were determined by specific enzyme kit for each parameter in Automatic System for Biochemistry. The global and differential blood cell count were performed at times 0, 12 and 24h after colostrum feeding. Colostrum has remained in the range of good quality according to the criteria of classification measures by colostrometer and digital Brix refractometer. Colostrum intake was the main factor of variation of the studied parameters. The evaluation of the passive transfer of immunity to newborn calves should be performed at about 24 hours after colostrum intake, because at this time, the absorption of colostral macromolecules is already finished and TP reaches stability. From this period, it is not possible to determine whether the assessed protein fractions are from colostrum or endogenous origin, which does not guarantee reliable assessment on the transfer of passive immunity.

Orientador: Nereu Carlos Prestes / Banca: José Paes de almeida Nogueira Pinto / Banca: Lisiane de Almeida Martins / Resumo: Diante da importância do colostro e leite de égua na alimentação e aquisição de imunidade de potros recém nascidos e na sua utilização na dieta humana, o objetivo do estudo foi avaliar a composição, propriedades físico-químicas, perfil microbiológico e contagem de células somáticas (CCS) do colostro e leite de éguas na fase inicial da lactação. As amostras foram coletadas de 36 éguas em sete momentos: M1= momento do parto, M2= 12 horas, M3= 24 horas, M4= 36 horas, M5= 48 horas, M6= onze dias e M7= 15 dias após o parto. Houve queda nos teores de proteína, gordura, sólidos totais e sólidos desengordurados, e aumento dos teores de lactose durante os momentos estudados. Também houve variação nos valores de pH, índice crioscópico e acidez durante os momentos, principalmente entre amostras de colostro e leite de transição. Houve isolamento microbiológico em 5,55% das amostras cultivadas, das quais 28,57% eram Streptococcus não hemolítico, 28,57% Streptococcus α hemolítico, 21,43% Streptococcus equi, 14,29% Micrococcus spp. e 7,14% Bacillus spp. Nenhum animal apresentou mastite clínica. Maiores valores de CCS foram encontrados no M1 e M2, com diminuição significativa no M3, com valores inferiores a 100.000 células somáticas/mL a partir deste momento. Perante os resultados podemos constatar alterações na composição, qualidade físico-química e celularidade durante os 15 primeiros dias de lactação. Onze amostras foram positivas ao California Mastitis Test, sendo observadas no M1 (36,36%), M2 (27,27%), M3 (18,19%), M5 (9,09%) e M7 (9,09%). Bactérias do gênero Streptococcus foram as mais frequentes isoladas. Entretanto, é necessário cautela na utilização de alguns métodos de diagnóstico de mastite, já que não há padronização dos mesmos para a espécie equina / Abstract: Given the importance of colostrum and milk of the mare on feeding and acquisition of immunity of newborn foals and their use in the human diet, the aim of the study was to evaluate the composition, physicochemical properties, microbiological profile and somatic cell count (SCC) of colostrum and milk of mares in early lactation. Samples were collected from 36 mares at seven moments: M1 = moment of partum, M2 = 12 hours, M3 = 24 hours, M4 = 36 hours, M5 = 48 hours, M6 = 11 days and M7 = 15 days post-partum. It was observed decreased levels of protein, fat, total solids and solids non-fat, and increased levels of lactose during the evaluated moments. There was also variation in pH, cryoscopic index and acidity during the moments, especially between samples of colostrum and transitional milk. The microbiological isolation occurred in 5.55% of the cultured samples, of which, 28.57% was non-hemolytic Streptococcus, 28.57% α-hemolytic Streptococcus, 21.43% Streptococcus equi, 14.29% Micrococcus spp. and 7.14% Bacillus spp. No animals showed clinical mastitis. Higher SCC values were found in M1 and M2, with a significant decrease in M3, with values less than 100,000 somatic cells / mL from this moment. Given these results, changes in the composition, physicochemical quality and cellularity during the first 15 days of lactation were observed. Eleven samples were positive on California Mastitis Test, being observed in M1 (36.36%), M2 (27.27%), M3 (18.19%), M5 (9.09%) and M7 (9.09%). Bacteria of the genera Streptococcus were the most frequently isolated. However, caution is needed in the use of some methods for diagnosis of mastitis, because there is no standardization in the equine species / Mestre